1. Field of the Invention
Embodiments of the invention described herein relate to antibodies directed to the antigen monocyte chemo-attractant protein-1 (MCP-1) and uses of such antibodies. In particular, in accordance with embodiments of the invention, there are provided fully human monoclonal antibodies directed to the antigen MCP-1. Nucleotide sequences encoding, and amino acid sequences comprising, heavy and light chain immunoglobulin molecules, particularly sequences corresponding to contiguous heavy and light chain sequences spanning the framework regions and/or complementarity determining regions (CDRs), specifically from FR1 through FR4 or CDR1 through CDR3, are provided. The antibodies of the invention find use as diagnostics and as treatments for diseases associated with the overproduction of MCP-1. Hybridomas or other cell lines expressing such immunoglobulin molecules and monoclonal antibodies are also provided.
2. Description of the Related Art
An increased production of angiogenic factors and decreased production of angiogenesis inhibitors by cancer cells, vascular endothelial cells and other stromal cell types are believed to induce tumor angiogenesis. Stroma, comprised of interstitial connective tissues, basal lamina, blood cells, blood vessels and fibroblastic cells, surround almost all solid tumor cells. Interactions between the stroma and cancer cells play a critical role in the neovascularization of tumors. Further, macrophage, which are also stromal components, are important in tumor angiogenesis. (M. Ono et al., Cancer Chemother. Pharmacol. (1999) 43(Suppl.): S69-S71.)
Macrophages are the major terminally differentiated cell type of the mononuclear phagocyte system, and are also one of the key angiogenic effector cells, producing a number of growth stimulators and inhibitors. A number of angiogenic cytokines are known to be produced by macrophages, including monocyte chemo-attractant protein 1 (MCP-1).
MCP-1 is known to be chemotactic for T lymphocytes, basophils and NK cells. MCP-1 is one of the most potent macrophage recruiting molecules. Once recruited to sites of inflammation or tumors, macrophages can generate a number of angiogenic cytokines, thereby stimulating pathologic angiogenesis. A number of studies have shown a relationship between angiogenesis, macrophage recruitment, and prognosis in patients with various kinds of tumors (G. Fantanini et al., Int. J. Cancer (1996) 67:615; N. Weidner et al., J. Natl. Cancer Inst. (1992) 84:1875). Leek et al. have further demonstrated that focally increased macrophage numbers are closely related to vascularization and prognosis in breast cancer patients (Cancer Res. (1996) 56:4625). R. Huang et al. (Cancer Res. (2002) 62:2806-2812) have shown that Connexin 43 suppresses human glioblastoma cell growth by down regulation of MCP-1, as discovered by using protein array technology.
Goede et al. (Int. J Cancer (1999) 82: 765-770) first demonstrated that MCP-1 had an angiogenic potency which was equivalent to that of VEGF when tested in a rabbit corneal model. In their model, the angiogenic activity induced by MCP-1 was associated with an intense recruitment of macrophages into the rabbit cornea. Salcedo et al. have reported that MCP-1 induced chemotaxis of human endothelial cells at nanomolar concentrations. This chemotactic response was inhibited by a polyclonal antibody to human MCP-1 (R. Salcedo et al., Blood (2000) 96(1):34-40).
MCP-1 is the predominant chemokine expressed in ovarian cancer (Negus, R. P. M. et al., J. Clin. Investig. (1995) 95: 2391-96; Sica, A. et al., J. Immunology (2000) 164(2):733-8). MCP-1 is also elevated in a number of other human cancers including bladder, breast, lung, and glioblastomas.
In addition, the importance of MCP-1 in inflammation has been shown in a number of studies. For example, H. J. Anders et al., have demonstrated chemokine and chemokine receptor expression during initiation and resolution of immune complex glomerulonephritis (J. Am. Soc. Nephrol. (2001) 12: 919-2001). Segerer et al. (J. Am. Soc. Nephrol. (2000) 11:2231-2242) also have studied the expression of MCP-1 and its receptor chemokine receptor 2 in human crescentic glomerulonephritis. J. A. Belperio et al. have shown a critical role for the chemokine MCP-1/CCR2 in the pathogenesis of bronchiolitis obliterans syndrome (J. Clin. Investig. (2001) 108: 547-556). N. G. Frangogiannis et al. have delineated the role of MCP-1 in the inflammatory response in myocardial infarction (Cardiovascular Res. (2002) 53: 31-47). Gerard and Rollins (Nature Immunol. (2001) 2:108-115) and Reape and Groot (Atherosclerosis (1999) 147: 213-225) have discussed the role of MCP-1 in atherosclerosis and other diseases. Also, Schmidt and Stern (Arterioscler. Thromb. Vasc. Biol. (2001) 21:297-299) describe MCP-1 interactions in restenosis.
Human MCP-1, a 76-amino-acid CC chemokine with an N-terminal pyroglutamic acid, was originally purified from several sources including phytohemagglutinin-stimulated human lymphocytes (Yoshimura, T. et al., J. Immunol. (1989) 142:1956-62), a human glioma cell line (Yoshimura, T., et al., J. Exp. Med. (1989) 169:1449-59), and the human myelomonocytic cell line THP-1 (Matsushima, K., et al., (1989) J. Exp. Med. (1989) 169: 1485-90). MCP-1 was first described as lymphocyte-derived chemotactic factor (LDCF). Other names for the protein are tumor-cell-derived chemotactic factor (TDCF), glioma-derived monocyte chemotactic factor (TDCF), glioma-derived monocyte chemotactic factor (GDCF), smooth muscle cell-derived chemotactic factor (SMC-CF), monocyte chemotactic activating factor (MCAF) and CCL2. Molecular cloning of the cDNA encoding MCP-1 (Furutani, Y., et al., (1989) Biochem. Biophys. Res. Comm. (1989) 169:249-55; B. J. Rollins, et al., Mol. Cell. Biol. (1989) 9:4687-95; Chang, H. C., et al., Int. Immunol. (1989) 1:388-97) revealed an open reading frame of 99 amino acids, including a signal peptide of 23 amino acids. The mouse homologue gene of MCP-1 was named JE (B. J. Rollins et al., 1989).
WO 200189565, published Nov. 29, 2001, discloses polyclonal antibodies to human MCP-1 and describes the inhibition of tumor growth in a nude mouse model by the use of such polyclonal antibodies.
Embodiments of the invention described herein relate to fully human monoclonal antibodies to human MCP-1 that block MCP-1-induced chemotaxis of THP-1 cells, a cell line derived from a patient with acute monocytic leukemia. These cells are used as a surrogate for assessing the migration of normal human mononuclear cells in circulation. Mononuclear cell infiltration stimulated by MCP-1 plays a pathologic role in a number of inflammatory conditions including rheumatoid arthritis, glomerulonephritis, atherosclerosis, transplant rejection, psoriasis, restenosis, and autoimmune diseases such as multiple sclerosis. An antibody that blocks MCP-1 activity and prevents monocyte infiltration will find use as a treatment for these and other inflammatory diseases.